Effects of reduced glutathione supplementation in semen freezing extender on frozen-thawed bull semen and in vitro fertilization

During cryopreservation, spermatozoa may suffer cold and cryo-induced injuries ―associated with alterations in cell defense systems― that are detrimental to their function and subsequent fertility. This study aimed to determine the efficacy of supplementing the semen freezing extender with the antioxidant reduced glutathione (GSH) in cattle. Semen was collected from four bulls and diluted in a freezing extender supplemented with or without GSH (0, 1, 5, and 10 mM) before the cooling step of the cryopreservation process. After thawing, the quality of the frozen-thawed semen was investigated for motility, viability, acrosomal and DNA integrity, and subsequent embryo development after in vitro fertilization of bovine oocytes. Additionally, semen from one of the bulls was used to analyze semen antioxidative potential, sperm penetration into oocytes, male pronucleus formation rate, and embryo DNA integrity. The sperm quality varied among bulls after GSH supplementation. One bull had decreased sperm total motility, and two bulls had decreased sperm DNA integrity. GSH supplementation had positive effects on embryo development for three bulls. Two of them showed both improved cleavage and blastocyst formation rates, while the other one only showed an improved cleavage rate. We observed positive effects on early male pronucleus formation and no negative effects on DNA integrity and cell number in blastocyst stage embryos. Although the effect varies depending on individual bulls and GSH concentration, GSH supplementation in semen may improve in vitro embryo production from frozen semen.     

Toe white line separation associated with the notch (crena marginalis) on the distal phalanges in Thoroughbred yearling horses

The shape of the white line of the hoof is closely related to the shape of the notch on the dorsal distal bearing border of the distal phalanx (P3). In this study, a radiographic survey of the P3 of both forelimbs of 163 Thoroughbred yearling horses was conducted. The correlation of the depth and width of the notch were analyzed with the toe white line separation grades (0 to 3). As a result, the toe white line separation grade increased, the depth and the width of the notch also increased significantly. Radiographic examination of the P3 of the forelimbs might be useful for deciding whether to implement hoof care to prevent onset of toe white line separation.     

A long-term observation for ecology of pathogenic Yersinia in wild rodents living in Fukushima Prefecture,Japan

From 2012 to 2021, prevalence of pathogenic Yersinia in wild rodents captured in Fukushima Prefecture, Japan was investigated twice a year to clarify the ecology of this pathogen in wild rodent populations. Pathogenic Yersinia enterocolitica O8 was isolated from 13 (1.7%) of 755 wild rodents. The Y. enterocolitica O8 isolates harbored three virulent genes (ail, fyuA, and virF). This pathogen was isolated repeatedly from wild rodents in April 2015, 2016, and 2017, in June and November 2020, and in April 2021, which was 6 of 19 times of observations. All Y. enterocolitica O8 isolates showed the same PFGE patterns. These results indicated that the same clone of pathogenic Y. enterocolitica O8 has been maintained in wild rodent populations in Fukushima Prefecture. Therefore, wild rodent populations contribute substantially to the continuous transmission of Y. enterocolitica O8 and its persistence in the ecosystem. This is the first report on the isolation of pathogenic Y. enterocolitica O8 in wild rodents in Fukushima Prefecture, Japan.     

Equine nonneoplastic abnormal ovary in a draft mare with high serum anti-Müllerian hormone: a case study

We performed a standing hand-assisted laparoscopic ovariectomy in a draft mare that presented with high serum anti-Müllerian hormone (AMH) level and had an enlarged single cystic ovary. Histopathological examination revealed no tumor cell proliferation in the ovary, but the presence of a large ovarian cyst was confirmed. In the diagnosis of abnormal ovaries in mares, a comprehensive assessment should be performed, including the monitoring of ovarian morphology and biomarkers over time, to determine the disease prognosis and treatment plan. The case of this mare with a nonneoplastic abnormal ovary and increased serum AMH level was rare. We suggest that standing hand-assisted laparoscopic ovariectomy is useful for the removal of large ovaries in draft mares.     

Evaluation of Low Level Laser Therapy on Primary Healing of Experimentally Induced Full Thickness Teat Wounds in Dairy Cattle

Objective —The purpose of this study was to evaluate the effect of low-level laser therapy (LLLT) on sutured wounds of the teat in dairy cattle.
Study Design —By using the Latin square design, the effect of LLLT was evaluated by radiography, measurement of microcirculation flow, histopathology, tensiometry, and hydroxyproline analysis.
Animals or Sample Population —Sixteen teats of four dairy cattle.
Methods —Full thickness wounds were made on the cranial surface of the teats. Teats were distributed into four groups; group A and B wounds were closed with a Gambee pattern, group C and D wounds were closed with three-layers of continuous suture pattern. Group B and D wounds were treated with 3.64 J/cm² of LLLT using a helium-neon system continuous wave (632.8 nm) output of 8.5 mW.
Results —The teat wall in non-LLLT groups was significantly thicker than in LLLT groups on day 7, 14 and 21. The mean blood flow differences between control and sutured sites in LLLT groups were significantly lower than those in non-LLLT groups. The morphology of the epidermis in LLLT groups more closely resembled the normal epidermis than that of non-LLLT groups. Collagen fibers in LLLT groups were denser, thicker, better arranged and more continuous with existing collagen fibers than those in non-LLLT groups. The mean tensile strength was significantly greater in LLLT groups than in non-LLLT groups.
Conclusion —The LLLT affects various aspects of the healing process, including minimizing inflammation, formation of edema, improvement of skin regeneration and enhancement of collagen synthesis.
Clinical Relevance —The LLLT could accelerate healing of sutured wounds of the teat in dairy cattle.     

Decrease in stearic acid proportions in adipose tissues and liver lipids in fatty liver of dairy cows

Samples of liver and perirenal, mesenteric and subcutaneous fat were collected from 16 sick necropsied dairy cows to evaluate the fatty acid profiles in the hepatic and adipose tissues associated with advanced fatty liver or hepatic lipidosis. Hepatic triglyceride and eight fatty acids were measured in the hepatic and adipose tissues. Six cows had more than 3% triglyceride on fresh weight in their livers and were classified as having fatty liver. Stearic and linoleic acid proportions in the liver decreased markedly with increased hepatic triglyceride levels, while the proportion of palmitic and oleic acids increased. The most striking fluctuations in hepatic lipidosis were manifested as decreased stearic acid in the adipose tissues including subcutaneous fat with the trend of decreasing stearic acid. Palmitic acid was elevated in hepatic and perirenal fat in fatty liver cows. In instances of advanced hepatic lipidosis, palmitoleic acid increased in only subcutaneous fat and not in perirenal or mesenteric fat. In addition to the proportions of hepatic fatty acids in fatty liver, this study also clarified the fluctuations observed in the profiles of fatty acids of the adipose tissues in cows with advanced hepatic lipidosis, particularly the decline in the proportions of stearic acid.     

The response of adipose tissues to Mycoplasma pulmonis and Sendai virus infection in C57BL/6 and DBA/2 mice

Adipose tissues in mammals are categorized into white and brown adipose tissues in which cellular morphology, cell functions, and tissue distribution are different. White adipose tissue (WAT) plays a major role in energy reservation, while brown adipose tissue (BAT) mainly relates to the thermoregulation of the body. One interesting function of adipose tissue is the response to the infection, especially the pathogens that cause pneumonia. We have previously reported that DBA/2 (D2) mice are susceptible to pathogens causing pneumonia, Mycoplasma (M.) pulmonis and Sendai virus (SeV), whereas C57BL/6 (B6) mice are resistant to them. Furthermore, morphological alteration of mediastinal fat tissue (MFT) was seen after infection of M. pulmonis in D2 mice but not in B6 mice. In this study, we aimed to exhibit the difference in adipose tissue response in other areas, including interscapular brown adipose tissue (iBAT), inguinal white adipose tissue (ingWAT), and perigonadal WAT (perigoWAT) between resistant strain, B6 and susceptible strain, D2 after challenging them with M. pulmonis and SeV. Compared with B6 mice, D2 mice showed an increase in fat-associated lymphoid cluster in MFT, an increase in BAT in both iBAT and ingWAT after M. pulmonis and SeV infection. The results of this study indicate that pneumonia caused by M. pulmonis and SeV infection induces browning of adipocyte, suggesting that BAT plays a role in pathogen infection and inflammation.